ABSTRACT
OBJECTIVE： To establish a method for simultaneous determination of 10 isoflavones in Belamcanda chinensis， and to evaluate the differences of active ingredient content of B. chinensis from different areas. METHODS： UPLC method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.5 ％ methyl-β-cyclodextrin and 0.1％ phosphate as water phase， acetonitrile as organic phase （gradient elution） at the flow rate of 0.2 mL/min. The column temperature was set at 35 ℃， and the detection wavelength was set at 265 nm. The sample size was 2 μL， and analysis time was 20 min. The contents of 10 isoflavones in 26 samples from 8 provinces， including tectoridin， iristectorin A， iristectorin B， iridin， tectorigenin， iristectorigenin B， iristectorigenin A， irigenin， irisflorentin， dichotomitin， were determined. RESULTS： The linear ranges of tectoridin， iristectorin A， iristectorin B， iridin， tectorigenin， iristectorigenin B， iristectorigenin A， irigenin， irisflorentin， dichotomitin were 8.569 5-342.78， 0.643-25.72， 1.119 8-44.79， 2.187 8-87.51， 0.770 3-30.81， 0.421 3- 16.85， 0.288 5-11.54， 1.795 3-71.81， 0.560 8-22.43， 0.086-3.44 μg/mL（all r≥0.999 6）. The limits of quantitation were 0.015， 0.102， 0.096， 0.013， 0.036， 0.088， 0.102， 0.019， 0.067， 0.092 μg/mL. RSDs of precision， stability（24 h）and reproducibility tests were lower than 2.00％ （n＝6）. The recoveries ranged 95.30％-103.30％ （all RSD≤2.33％， n＝6）. Among 26 samples of B. chinensis， the content of tectoridin was the highest （3.66％-57.79％）， and the content of dichotomitin was the lowest （0.09％- 0.59％）， the contents of irisflorentin were 0.29-2.80 mg/g. The contents of isoflavones in B. chinensis from different areas were different greatly. CONCLUSIONS： The established method is sensitive， with short analysis time and good repeatability， and can be used to determine the content of 10 isoflavones and evaluate the content difference of each component.